甘油氧化酶法测定血清三酰甘油的研究

目的:建立一种甘油氧化酶法测定血清三酰甘油新法。方法:用脂蛋白脂酶、甘油氧化酶和3-甲基-N-乙基(β-羟乙基)苯胺(简称MEHA)色原系统进行血清三酰甘油测定。结果:对最适缓冲液浓度、pH和种类的选择,提高了灵敏度,线性范围为0～18 mmol/L,正常参考范围为男0.05～1.80 mmol/L、女0.04～1.70 mmol/L。其回收率为93%～103%,精密度:批内(n=20)CV=2.6%～4.2%,批间(n=20)CV=3.1%～4.8%,与酶(GPO-PAP)法比较,相关性良好(γ=0.982)。结论:本法具有高度的特异性和准确性,操作简便、灵敏快速、标本用量少、结果稳定可靠,是一种新的三酰甘油测定法。     

噻奈普汀钠片溶出度测定

目的　建立噻奈普汀钠片溶出度测定方法。方法　以水为溶出介质 ,转篮法 ,转速 10 0r/min ,4 5min为取样点 ,检测波长为 2 2 5nm。结果　噻奈普汀钠在 2 .74～ 2 7.4 4 μg·ml-1范围内 ,浓度与吸收度呈良好线性关系 (r =0 .9998)。回收率为 10 0 .1% ,RSD 为 0 .6 1% ,三批溶出度平均值均大于 90 %。结论　本法快速简便 ,为噻奈普汀钠片体外溶出监测提供了方法     

血清LDH紫外监测法试剂盒的研制和应用

作者研制了适用于在自动生化仪上测定血清LDH活性的LD-P紫外监测法试剂盒.实验室研究证明,自制试剂盒在Impact 400型自动生化仪上,与西德BM公司,美国Gilford公司同类试剂盒对比测定结果一致,呈非常显著相关.酶促反应的延迟时间短(<20s),血清LDH活性测定线性范围宽,可达1000u/L(37℃),批内变异系数1.1～4.7%.在4～8℃条件下贮存,本试剂盒可稳定7个月,重溶试剂可稳定7天.81名健康成人血清LDH活性测定结果为318.3±61.9u/L(37℃).     

薄层扫描法测定脑康灵口服液中黄芪甲苷和人参皂苷Re的含量

目的　建立脑康灵口服液的含量测定方法。方法　展开剂为正丁醇 -乙酸异戊酯 -水 (4∶1∶5 )溶液 ,显色剂为 2 0 %硫酸乙醇溶液 ,采用双波长薄层扫描法 ,对脑康灵口服液中黄芪和西洋参的有效成分黄芪甲苷及人参皂苷Re同时进行含量测定。结果　黄芪甲苷在 0 .4 9～ 2 .94 μg范围内呈良好的线性关系 ,r=0 .9996 ,平均回收率 97.93% ,RSD 为 0 .90 % ;人参皂苷Re在 1.0 6～ 6 .36 μg范围内呈良好的线性关系 ,r =0 .9987,平均回收率 98.2 4 % ,RSD为 1.0 4 % ,5批样品中黄芪甲苷和人参皂苷Re的含量平均值为 0 .0 383、0 .32 70mg·ml-1。结论　该方法简便、准确 ,重现性好 ,可用于该制剂的含量测定及质量控制。     

有机磷化合物体外全血胆碱酯酶检测方法学研究

目的建立一种在体外快速测定乙酰胆碱酯酶（AChE）活性的方法,为有机磷毒剂中毒的诊断及救治提供依据。方法用改良的5,5-连硫代-双-2-硝基苯甲酸（DTNB）法测定AChE活性,以兔全血作为酶源进行方法学研究,并用不同种属动物全血进行了适用性考察,用SPECTRA MAX 190读板机测定412 nm处光密度的变化即可测定AChE活性。结果不同动物全血40倍稀释后作为酶源较为合适,批内差异为0.44%～0.67%,批间差异为1.03%～2.27%,该方法适用于兔、比格犬、豚鼠和猴全血AChE测定。梭曼和沙林对兔全血AChE活性抑制具有明显的量效关系。结论该方法简便、易行,适用于不同种属动物全血AChE活性的测定。     

血清ALT紫外监测法试剂盒的研制和应用

作者采用国产原料研制成功适用于在临床生化自动分析仪或紫外分光光度计上测定血清ALT(GPT)活性的ALT紫外监测法试剂盒.在Impact400型生化仪上所作的试剂盒性能质量评价实验证明,酶促反应的延迟时间短(<20s),批内变异系数1.1～6.5%,血清ALT活性线性可达1000u/L(37℃),与进口同类试剂盒对比测定结果一致,呈非常显著相关.自制ALT试剂盒4～8℃存放可稳定6个月.81名健康成人血清ALT活性为26.91±8.91u/L(37℃).     

双波长分光光度法测定蜂蜜普鲁卡因合剂中盐酸普鲁卡因及尼泊金乙酯的含量

本文采用双波长分光光度法测定蜂蜜普鲁卡因合剂中盐酸普鲁卡因及尼泊金乙酯的含量.盐酸普鲁卡因在3～15μg/ml范围内与吸收度呈良好的线性关系(r=1.0000),回收率为99.88%、RSD为0.23%;尼泊金乙酯在1.2～6.0μg/ml范围内与吸收度呈良好的线性关系(r=0.9999),回收率为99.85%,RSD为0.40%.该法操作简单,快速、准确.适应于医院对该制剂的质量控制.     

离子对萃取和高效液相色谱-电化学检测法测定血浆中儿茶酚胺

目的 :建立简便、灵敏的测定血浆儿茶酚胺的方法。方法 :利用离子对萃取法提取血浆中的儿茶酚胺 ,用高效液相色谱 电化学检测法 (HPLC ECD)进行检测 ,测定其回收率、线性范围、精密度及最低检测限 ,并测定和比较了大鼠和小鼠血浆中儿茶酚胺浓度。结果 :肾上腺素 (E)、去甲肾上腺素 (NE)、多巴胺 (DA)的加样回收率分别为(10 3.2± 4 .9) % ,(10 6 .1± 6 .3) %和 (10 1.1± 9.0 ) %。日内和日间精密度分别小于 14 .4 %和 11.6 %。该方法测定E和NE在 0 .5～ 16ng范围内 ,DA在 0 .2 5～ 8ng范围内含量与峰高比之间呈良好的线性关系 ,相关系数为 0 .9980～0 .9999,最低检测限分别为 :E 0 .2 5ng ,NE 0 .30ng,DA 0 .2 0ng。大鼠血浆中E浓度明显高于小鼠 ,NE和DA浓度无明显差异。结论 :本法用于检测血浆中儿茶酚胺水平具有简便、灵敏、快速、准确的特点 ,适合于小动物血浆中儿茶酚胺浓度的测定。     

分光光度法测定阿奇霉素注射液的含量

目的　建立分光光度法测定阿奇霉素注射液的含量。方法　精取阿奇霉素注射液适量 ,加 0 .1mol·L-1盐酸溶解并稀释至一定浓度 ,加入 85 %硫酸 ,涡旋混合 ,静置显色 ;在 4 82nm波长处按拟定方法测定。结果　浓度在 2 0～ 70mg·L-1范围内呈线性关系 ,回归方程为 :A =0 .0 0 932 C+0 .0 2 18,r=0 .9999;平均回收率 10 1.33% (n=9) ,日内、日间精密度 RSD<1%。结论　本法准确、简便、快速 ,可用于阿奇霉素注射液的质量控制。     

紫外分光光度法测定苄索氯铵插层蒙脱土中苄索氯铵的含量

目的:建立一种快速测定苄索氯铵含量的方法以及测定该紫外分光光度计对苄索氯铵的最小浓度.方法:采用紫外分光光度法在269nm波长处测定苄索氯铵含量.结果:苄索氯铵的含量线性范围0.05～0.3 mg/ml线性回归方程:A=2.8005C 0.0332 r=0.9997,平均回收率为100.01%,RSD=0.11%.结论:本操作简便、快速,结果准确.     

Spectrophotometric method for determination of bifunctional macrocyclic ligands in macrocyclic ligand-protein conjugates

A simple spectrophotometric assay for determination of bifunctional polyazacarboxylate-macrocyclic ligands of different sizes that are conjugated to proteins has been developed for: 12-membered macrocycle DOTA (2-[4-nitrobenzyl]-1, 4, 7, 10-tetraazacyclododecane-N,N′,N″,N-tetraacetic acid) and analogs, the 15-membered PEPA macrocycle (2-[4-nitrobenzyl]-1,4,7,10,13-pentaazacyclopentadecane-N,N′,N″,N,N-pentaacetic acid), and the large 18-membered macrocycle HEHA (1,4,7,10,13,16-hexaazacyclooctadecane-N,N′,N″,N,N,N′-hexaacetic acid). The method is based on titration of the blue-colored 1:1 Pb(II)-Arsenazo III (AAIII) complex with the polyazacarboxylate macrocyclic ligand in the concentration range of 0–2.5 μM, wherein color change occurring upon transchelation of the Pb(II) from the AAIII to the polyazamacrocyclic ligand is monitored at 656 nm. The assay is performed at ambient temperature within 20 min without any interfering interaction between the protein and Pb(II)–AA(III) complex. Thus, this method also provides a ligand-to-protein ratio (L/P ratio) that reflects the effective number of ligands per protein molecule available to radiolabeling. The method is not suitable for 14-membered TETA macrocycle (2-[4-nitrobenzyl]-1, 4, 8, 11-tetraazacyclotetradecane N,N′,N″,N-tetraacetic acid) because of low stability constant of Pb(II)-TETA complex. The method is rapid, simple and may be customized for other polyazacarboxylate macrocyclic ligands.     

Methylene blue-enhanced stability of (99mTc)HMPAO and simplified quality control--a comparative investigation.

(99mTc)HMPAO is a radiopharmaceutical used for SPECT imaging of regional cerebral perfusion and for labeling cellular blood elements. The addition of methylene blue enhanced the stability of lipophilic (99mTc)HMPAO complex up to 3 h after reconstitution, 86.9+/-4.2% compared to 49.8+/-8.9% for the non-stabilised complex and persists over time (86.2+/-3.5% after 15 min compared to 78.2+/-4.0% after 3 h). The method widely used for estimation of radiochemical purity is a standard chromatographic procedure which is quite time-consuming taking about 30 min. Comparison of the more rapid and simple solvent extraction method with octanol, which needs only about 10 min to complete. showed a good correlation with the chromatographic method (84.4+/-4.4% compared to 89.1+/-4.3%). Using ethyl acetate as solvent instead of octanol gave a slightly higher extraction rate of the lipophilic complex (91.5+/-5.5% compared to 89.1+/-4.3%). Further, the ethyl acetate extraction method results in an overestimation, extracting partly secondary complex. It is confirmed, that the stability of the lipophilic (99mTc)HMPAO complex can be increased with methylene blue. The octanol extraction method, using higher extraction volume (up to 3 ml), is recommended as a fast and efficient approach for the quality control in daily clinical routine.     

Simple biodosimetry method for use in cases of high-dose radiation exposure that scores the chromosome number of Giemsa-stained drug-induced prematurely condensed chromosomes (PCC)

There is a need for quick dose estimation by a simple method in radiation accidents. This study develops a simple and rapid dose estimation protocol for victims of such accidents, in particular those involving high radiation doses. Human peripheral blood lymphocytes (PBL) were gamma-irradiated in vitro at several dose points up to 60 Gy, and were stimulated with phytohaemagglutinin-P (PHA-P) for 2 days to obtain dividing cells. PBL were then forced to condense prematurely, using 50 nM calyculin A, and the obtained chromosome spreads were Giemsa stained. The G2-PCC (prematurely condensed chromosomes) index and chromosome number for each radiation dose point were scored. G2-PCC were stably induced using calyculin A within 24 h delays in stimulation of PBL with PHA-P. The chromosome number of G2-PCC increased steeply with radiation doses up to 30 Gy at a rate of 0.31 Gy(-1) and then decreased at 0.30 Gy(-1) up to 40 Gy. More than 10% of G2-PCC index remained up to a 15 Gy dose. Even after 40 Gy irradiation, about 2% PCC index was obtained, and this value was enough to score a sufficient number of chromosome spreads for analysis. Therefore, the combined use of chromosome number and G2-PCC index allows biodosimetry to be done easily and rapidly. If PCC are not induced using calyculin A, it is strongly suggested that the radiation dose is over 50 Gy. A rapid and easy dose estimation for large dose exposure whole-body was realized by combined analysis of Giemsa-stained chromosome number of G2-PCC and PCC index using calyculin A. This simple method will be of use for rapid decision making of therapy for radiation accident victims. This method also has potential for use as a biodosimeter for partial-body exposure accidents.     

Color vision with rapid-onset acceleration

INTRODUCTION: Only sporadic information exists concerning perceived color shifts at increased G-loads. The purpose of this study was to investigate whether or not color vision is affected by rapid onset high G7-loads up to +9 Gz, and specifically whether perception of hue changes. METHODS: There were 10 male subjects, 9 with normal color vision and 1 with red-green protanomaly, all accustomed to Gz-loads in a human centrifuge. Each subject was tested on a total of 60 Gz-exposures with 10 s periods at +3, +5, +7, and +9 Gz in the centrifuge on three different days. G-onset rate was 6 G x s(-1). The subjects wore an anti-G suit and performed straining maneuvers if necessary to maintain vision. Five square color stimuli of medium saturation (yellow, red, blue, green, and gray) were projected one at a time on a screen in front of the subject, who gave his hue response orally. RESULTS: In 96.6% of exposures to various Gz-loads, the subjects responded by correctly naming colors. (The statistical analyses of the results were done for the subjects with normal color vision, with the protanomalous subject excluded.) Hue shifts occurred at the higher +Gz-levels, including 7.7% of the +9 Gz exposures. Yellow was the hue most frequently perceived as changed. Hue shifts were reported for yellow in 11% and 16% of the +7 and +9 Gz exposures, respectively. Hue shifts at +9 Gz occurred as frequently as blackout and G-LOC together. However, statistical analyses showed no significant effects for +Gz-load. CONCLUSIONS: Absolute identification of the color stimuli of medium saturation was stable and was not significantly affected by the rapid onset +Gz-loads up to and including +9 Gz.     

A real-time smartphone-enabled time since death estimation from vitreous humour protein

This study reports on a smartphone-based application for the determination of time since death (TSD) from vitreous humour (VH) protein via recognition and quantification using a simple biuret method for on-the-spot analysis. In basic pH, in the presence of protein, the biuret reagent forms a protein-biuret complex resulting in a colour change that is measurable using a UV-vis spectrophotometer. The limit of detection (LOD) for Bovine serum albumin (BSA) was found to be 8 μg/ml and the limit of quantification (LOQ) was found to be 10 μg/ml. The developed method was further validated and applied for TSD determination from VH samples in the range of 3–60 h. The results showed that there is a linear correlation between the VH protein concentration and TSD, as the protein concentration decreases up to 60 h, with the equation TSD = 33.72 + (–0.27) × concentration. Further, in the present study smart approach was applied for the TSD determination. The results obtained showed that the current method proves to have a great utility up to 60 h. The linear regression equation between TSD and RGB (red, green and blue) intensities of protein concentration tup to 60 h ± 4.5 h was found to be TSD = 6.27E2+1.41*RGB intensity. The proposed method provides a smart, rapid and sensitive, as well as cost effective, TSD determination via VH protein.     

Simple biodosimetry method for use in cases of high-dose radiation exposure that scores the chromosome number of Giemsa-stained drug-induced prematurely condensed chromosomes (PCC)

There is a need for quick dose estimation by a simple method in radiation accidents. This study develops a simple and rapid dose estimation protocol for victims of such accidents, in particular those involving high radiation doses. Human peripheral blood lymphocytes (PBL) were γ-irradiated in vitro at several dose points up to 60?Gy, and were stimulated with phytohaemagglutinin-P (PHA-P) for 2 days to obtain dividing cells. PBL were then forced to condense prematurely, using 50 nM calyculin A, and the obtained chromosome spreads were Giemsa stained. The G2-PCC (prematurely condensed chromosomes) index and chromosome number for each radiation dose point were scored. G2-PCC were stably induced using calyculin A within 24?h delays in stimulation of PBL with PHA-P. The chromosome number of G2-PCC increased steeply with radiation doses up to 30?Gy at a rate of 0.31?Gy^???1 and then decreased at 0.30?Gy^???1 up to 40?Gy. More than 10% of G2-PCC index remained up to a 15?Gy dose. Even after 40?Gy irradiation, about 2% PCC index was obtained, and this value was enough to score a sufficient number of chromosome spreads for analysis. Therefore, the combined use of chromosome number and G2-PCC index allows biodosimetry to be done easily and rapidly. If PCC are not induced using calyculin A, it is strongly suggested that the radiation dose is over 50?Gy. A rapid and easy dose estimation for large dose exposure whole-body was realized by combined analysis of Giemsa-stained chromosome number of G2-PCC and PCC index using calyculin A. This simple method will be of use for rapid decision making of therapy for radiation accident victims. This method also has potential for use as a biodosimeter for partial-body exposure accidents.     

Spectrophotometric determination of Sn(II) using palladium chloride in kits for 99mTc radiopharmaceuticals.

A method of quantitating Sn(II) suitable for the analysis of kits for 99mTc radiopharmaceuticals, based on a spectrophotometric determination using the Pd(II)-Sn(II) complex (yellow species) is described. The absorbance of the complex is measured at 410 nm and Beer's law is obeyed up to 250 micrograms Sn(II) in the aqueous phase. Several radiopharmaceutical kits were analyzed for their Sn(II) content. The investigation indicates that the procedure is simple, rapid and accurate for quantitative estimation of Sn(II) in various 99mTc-labelling kits during development, manufacture and storage.     

Coronary artery bypass graft imaging using ECG-gated multislice computed tomography: comparison with catheter angiography

AIM: To compare the value of multislice computerized tomography (MSCT) in imaging coronary artery bypass grafts (CABGs) by direct quantitative comparison with standard invasive angiography. METHODS: Using MSCT, 50 consecutive patients who had previously undergone CABG surgery and had recently undergone invasive angiography for recurrent angina pectoris, were studied further using MSCT after intravenous injection of non-ionic contrast agent; cardiac imaging was performed during a single breath-hold. Graft anatomy was quantified, using both quantitative coronary angiography (QCA) and MSCT, by different investigators blinded to each other. Reproducibility was quantified using the standard error of the measurement expressed as a percentage in log-transformed values (CV%) and intraclass correlation (ICC). RESULTS: All 150 grafts were imaged using MSCT; only 4 patent grafts were not imaged using selective angiography. Good agreement was achieved between MSCT and QCA on assessment of proximal anastomoses (CV% 25.2, ICC 0.84), mid-vessel luminal diameter (CV% 15.5, ICC 0.91) and aneurysmal dilations (CV% 14.3). Reasonable agreement was reached on assessment of distal anastomoses (CV% 26.7, ICC 0.66) and categorization of distal run-off (ICC 0.73). Good agreement was observed for stenoses of over 50% luminal loss (CV% 8.7, ICC 0.97) but agreement on assessment of less severe lesions was poor (CV% 208.7, ICC 0.51). CONCLUSION: This study demonstrates that CABGs can be quantitatively evaluated using MSCT, and that significant lesions present in all CABG segments can be reliably identified. Agreement between MSCT and QCA for lesions of less than 50% luminal loss was poor.     

Simple and rapid determination of myristicin in human serum

Myristicin (5-allyl-1-methoxy-2,3-methylenodioxybenzene) is the main component of nutmeg (Myristica fragrans Houtt.) essential oil. The increasing use of myristicin as a cheap hallucinogenic intoxicant, frequently causing fatal cases of myristicin poisoning, requires new methods for determination of this compound in blood. This report describes the rapid, simple, and useful procedure for myristicin analysis in human serum, involving myristicin–protein complex degradation before chromatographic analysis. The developed method is characterized by a high recovery (above 99 %), a low detection limit (6.0 ng/g) and good repeatability (average RDS of 2.01 %).     

复方乳酸钠注射液中乳酸钠含量测定方法的比较研究

目的：建立一种重现性好,操作相对简单的乳酸钠含量测定方法。方法：以离子交换法（药典法）作对照,进行氧化还原法,中和法和紫外分光光度法的回收率比较研究。结果：氧化还原法和中和法与离子交换法无显著性差异（P〉0.05）,紫外分光光度法与离子交换法有显著性差异（P〈0.05）。结论：药典法结果准确、重现性好,但操作繁琐;中和法操作简单快速,但终点不易观察;紫外分光光度法简便易行,但偏差较大。改进后的氧化还原法操作简便,结果准确可靠,重现性好,适合医院制剂的分析要求,是乳酸钠含量测定比较理想的方法。